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牛精子的形态缺陷、精子 DNA 完整性和顶体蛋白化。

Morphological defects, sperm DNA integrity, and protamination of bovine spermatozoa.

机构信息

School of Veterinary Science, The University of Queensland, Gatton, QLD, Australia.

School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD, Australia.

出版信息

Andrology. 2018 Jul;6(4):627-633. doi: 10.1111/andr.12486. Epub 2018 Apr 6.

Abstract

The association between sperm morphology characteristics and DNA conformation and integrity is still controversial. In bulls, major morphological sperm abnormalities have been associated with reduced fertility, and morphological assessment is used to provide an indication of potential fertility of the individual. Sperm DNA fragmentation and damage has a negative effect on embryo development and subsequently fertility, with bull spermatozoa generally displaying low levels of DNA damage and tight chromatin. However, sensitive methods for detecting chromatin damage may reveal associations with morphological defects. The objective was to determine whether morphological sperm abnormalities and variables expressing sperm DNA integrity and protamination are correlated in bulls, using the sperm chromatin structure assay (SCSA) and the sperm protamine deficiency assay (SPDA). Electroejaculated samples (n = 1009) from two-year-old tropically adapted bulls were split and fixed and submitted to microscopic sperm morphology assessment, and snap-frozen for sperm nuclear integrity assessments by SPDA and SCSA. For SPDA, the variables were defective (MCB) and deprotaminated (HCB), and for SCSA, the variables were DNA fragmentation index (DFI) and high DNA stainability (HDS). HCB correlated with DFI; τKen = 0.317 and HDS; 0.098, and MCB correlated with DFI; 0.183 (p < 0.001). The percentage of morphological normal spermatozoa was correlated negatively to DFI; τKen = -0.168, MCB; -0.116 and HCB; -0.137 (p < 0.001). HCB and DFI were both positively correlated to head defects, proximal droplets, and spermatogenic immaturity, but not to distal droplets, vacuoles, or diadems. Sperm DNA integrity and protamination, using the SCSA and SPDA, respectively, in bulls show associations with morphological parameters, particularly with head shape abnormalities and indicators of spermatogenic immaturity, including proximal droplets. The vacuoles and diadem defects were not correlated with sperm nuclear integrity, and hence, these are likely physiological features that may not directly affect sperm chromatin configuration.

摘要

精子形态特征与 DNA 构象和完整性之间的关系仍存在争议。在公牛中,主要的形态学精子异常与生育力降低有关,形态学评估用于提供个体潜在生育力的指示。精子 DNA 碎片化和损伤对胚胎发育和随后的生育力有负面影响,公牛精子通常显示出低水平的 DNA 损伤和紧密的染色质。然而,用于检测染色质损伤的敏感方法可能会揭示与形态缺陷的关联。本研究的目的是使用精子染色质结构分析(SCSA)和精子鱼精蛋白缺乏分析(SPDA)来确定公牛的形态学精子异常和表达精子 DNA 完整性和鱼精蛋白的变量是否相关。从两岁的热带适应公牛采集电刺激精液样本(n=1009),并将其分为两部分,一部分固定后进行显微镜下精子形态评估,另一部分用于通过 SPDA 和 SCSA 进行精子核完整性评估的快速冷冻。对于 SPDA,变量是缺陷(MCB)和去鱼精蛋白(HCB),对于 SCSA,变量是 DNA 碎片化指数(DFI)和高 DNA 染色性(HDS)。HCB 与 DFI 相关;τKen=0.317 和 HDS;0.098,MCB 与 DFI 相关;0.183(p<0.001)。形态正常精子的百分比与 DFI 呈负相关;τKen=-0.168,MCB;-0.116 和 HCB;-0.137(p<0.001)。HCB 和 DFI 均与头部缺陷、近端滴状液和精子发生不成熟呈正相关,但与远端滴状液、空泡或冠部无关。使用 SCSA 和 SPDA 分别在公牛中进行精子 DNA 完整性和鱼精蛋白分析,显示与形态参数相关,特别是与头部形状异常和精子发生不成熟的指标相关,包括近端滴状液。空泡和冠部缺陷与精子核完整性不相关,因此,这些可能是生理特征,不会直接影响精子染色质构型。

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